Impact of the Epigenetically Regulated Hoxa-5 Gene in Neural Differentiation from Human Adipose-Derived Stem Cells Hernández, Rosa Jiménez Luna, Cristina Prados Salazar, José Carlos Ortiz Quesada, Raúl Perazzoli, Gloria Melguizo Alonso, Consolación Mesenchymal Stem Cells Neuronal differentiation Epigenetic changes Hoxa-5 CRISPR/dCas9 Human adipose-derived mesenchymal stem cells (hASCs) may be used in some nervous system pathologies, although obtaining an adequate degree of neuronal differentiation is an important barrier to their applicability. This requires a deep understanding of the expression and epigenetic changes of the most important genes involved in their differentiation. We used hASCs from human lipoaspirates to induce neuronal-like cells through three protocols (Neu1, 2, and 3), determined the degree of neuronal differentiation using specific biomarkers in culture cells and neurospheres, and analyzed epigenetic changes of genes involved in this differentiation. Furthermore, we selected the Hoxa-5 gene to determine its potential to improve neuronal differentiation. Our results showed that an excellent hASC neuronal differentiation process using Neu1 which efficiently modulated NES, CHAT, SNAP25, or SCN9A neuronal marker expression. In addition, epigenetic studies showed relevant changes in Hoxa-5, GRM4, FGFR1, RTEL1, METRN, and PAX9 genes. Functional studies of the Hoxa-5 gene using CRISPR/dCas9 and lentiviral systems showed that its overexpression induced hASCs neuronal differentiation that was accelerated with the exposure to Neu1. These results suggest that Hoxa-5 is an essential gene in hASCs neuronal differentiation and therefore, a potential candidate for the development of cell therapy strategies in neurological disorders. 2021-09-22T09:52:42Z 2021-09-22T09:52:42Z 2021 info:eu-repo/semantics/article Hernández, R.; Jiménez-Luna, C.; Ortiz, R.; Setién, F.; López, M.; Perazzoli, G.; Esteller, M.; Berdasco, M.; Prados, J.; Melguizo, C. Impact of the Epigenetically Regulated Hoxa-5 Gene in Neural Differentiation from Human Adipose-Derived Stem Cells. Biology 2021, 10, 802. https://doi.org/ 10.3390/biology10080802 http://hdl.handle.net/10481/70353 10.3390/biology10080802 eng http://creativecommons.org/licenses/by/3.0/es/ info:eu-repo/semantics/openAccess Atribución 3.0 España MDPI